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. 2020 Aug 10;9:e57525. doi: 10.7554/eLife.57525

Figure 6. UPF3B shapes olfactory neurogenesis.

(A) The fraction of HBCs per all OSN precursors/OSNs (HBCs, GBCs, iOSNs and mOSNs) (left) or all OE cells (right), in Upf3b-null (KO) and WT mice, as determined by scRNA-seq analysis. *, p<0.05. (B) The percentage of cells from the indicated cell sub-clusters in Upf3b-null (KO) and WT mice, as determined by scRNAseq analysis. (C) Cell number in each cell sub-cluster, as defined in Figure 3D. (D) Most statistically enriched GO terms in the mOSN-2 and −4 sub-clusters. (E) Heatmap depicting the expression pattern of anti-microbial genes in the indicated cell subsets. (F) Left: Western blot analysis of endogenous CAMP protein level in the OE from Upf3b-null (KO) and WT mice. Right: quantification of CAMP level normalized against GAPDH (n = 3). *, p<0.05. (G) IF analysis of adult mouse OE sections co-stained with antisera against CAMP (red) and OMP (green). Nuclei were stained with DAPI (blue). (H, I) The percentage of mOSNs (H) and iOSNs (I) in our scRNAseq datasets that express Olfr genes. Left, all known Olfr genes. Right, the 78 Olfr genes significantly downregulated in Upf3b-null mice, based on RNAseq analysis (Figure 1C). *, p<0.05.

Figure 6.

Figure 6—figure supplement 1. Impact of UPF3B loss on OE cell subsets.

Figure 6—figure supplement 1.

(A) Left, IF analysis of adult mouse OE sections co-stained with antisera against TRP63 (red) and OMP (green). White arrows label TRP63+ HBCs. (n = 3). Right, quantification of TRP63+ cells per 1 mM. For each group, 15 different OSN field lengths (75 μm long) from three individual mice were examined to calculate the number of TRP63+ cells per mM. (B) The fraction of different cell subsets per OSN precursors and OSNs (HBCs, GBCs, iOSNs and mOSNs) cells (top) or all OE cells (bottom), in indivdiual Upf3b-null (KO) and control (WT) mice, as determined by scRNA-seq analysis. (C) Left, western blot analysis of OMP protein expression in independent OE samples from Upf3b-null and control (wild type) mice. GAPDH is the normalization control. Right, quantification of OMP expression normalized to GAPH levels (n = 4).
Figure 6—figure supplement 2. mOSN subsets and UPF3B-regulated genes.

Figure 6—figure supplement 2.

(A) Scatter plots showing Pearson correlation analysis of the transcriptomes from the indicated mOSN cell sub-clusters. (B) Left, IF analysis of adult mouse OE sections co-stained with antisera against CAMP/CRAMP (red) and OMP (green). Nuclei were stained with DAPI (blue). (n = 3). Right, quantification of OMP+ cells per mm2. For each group, 6 different mSONs views (2500 μm2) from 3 individual mice were quantified to calculate cell number per mm2. (C) The chromosomal distribution of the 78 Upf3b-regulated Olfr genes.